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Expression and characterization of the preS1 peptide of hepatitis B surface antigen in Escherichia coli
Author(s) -
Lin Yuan,
Liu YanXin,
Cislo Teresa,
Mason Bobby L.,
Yu MeiYing W.
Publication year - 1991
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890330308
Subject(s) - recombinant dna , fusion protein , microbiology and biotechnology , antigen , chemistry , hepatitis b virus , peptide , open reading frame , plasmid , escherichia coli , peptide sequence , gene , virology , biology , virus , biochemistry , genetics
The infectious particles of hepatitis B virus (HBV) contain 3 related surface antigens, i.e., small, medium, and large, all of which are encoded by one large open reading frame with multiple initiation codons. The large surface antigen (L‐Ag) contains preS1, preS2, and S regions while both the middle and small surface antigens lack preS1. Several lines of evidence suggested that the preS1 region is involved in the binding of HBV to human hepatocytes as shown by its binding to HepG2 cells and isolated human he‐patocyte membranes. To obtain large quantity of preS1 peptide, an expression vector was constructed containing a lac promoter, the 5′ half of the p‐galactosidase gene, the Factor Xa tetrapep‐tide recognition sequence, and the coding region of preS1 plus preS2. This recombinant plasmid constitutively produced high concentration of a fusion protein in inclusion bodies in Escherichia coli. When the fusion protein was treated with Factor Xa, a peptide consisting of the N‐terminal 91 amino acids of the preS1 region was released. This preS1 fragment purified by anion exchange chromatography was able to bind specifically to the isolated plasma membranes from human liver. Hence, this recombinant preS1 peptide can be used to identify and isolate hepatocyte receptors for HBV.